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Epigenetic mechanism in search for the pathomechanism of diabetic neuropathy development in diabetes mellitus type 1 (T1DM)

  • Endocrine Genetics/Epigenetics
  • Published:
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Abstract

Objective

The aim of this study was to check the hypothesis concerning the crucial role of DNA methylation (one of the epigenetic mechanisms) within selected genes related to the destruction and regeneration of neural cells and its input in the pathogenesis of diabetic neuropathy, using a model of the DNA in peripheral blood cells.

Methods

A cross-sectional, case-control study was conducted, consisting of 24 adult Type 1 Diabetes Melitus (T1DM) patients with autonomic neuropathy (CAN), 25 T1DM patients without neuropathy and 25 matched, healthy adults acting as a control (Ctrl). The Ewing’s tests, using the ProSciCard apparatus (Mewicon CATEEM-Tec GmbH), was employed to assess the severity of the patients’ symptoms of autonomic neuropathy. For DNA methylation analysis, DNA material of each sample DNA after bisulfite conversion was used for the hybridization of BeadChips (Infinium Methylation EPIC Kit, Illumina), and imaged on the Illumina HiScan. The changes in the expression of selected genes were examined using real-time PCR. Probes were labeled using fluorescein amidite, FAM (Thermo Fisher Scientific). Amplification was performed using the continuous fluorescence detection 7900 HT Fast Real-Time PCR system (Thermo Fisher Scientific). The expression ratio of the target mRNA was normalized to the level of 18s RNA and compared with the control. Statistical analysis was performed using Statistica version 13.1. The statistically significant results were recognized, with a value of p < 0.05.

Results

Clinical analysis of the investigated groups revealed a significantly higher percentage of personal insulin pump users in the group without neuropathy. The glucose metabolic control, based on the HbA1c level analysis, was also significantly better in T1DM patients without CAN. The Bumphunter method for DNA methylation analysis showed statistically significant regions related to the genes involved in nerve regeneration ninjurin 2 (NINJ2) and functionality (BR serine/threonine kinase 2 BRSK2, claudin 4 CLDN4). When compared with T1DM patients without neuropathy, T1DM patients with neuropathy showed significantly increased methylation in the first NINJ2 axon, and a lower level of DNA methylation in the region of the first intron of BRSK2, as well as the CLDN4 5′UTR regions. The qRT-PCR results confirmed the decreased expression of NINJ2 and CLDN4 genes in patients with T1DM with CAN.

Conclusions

The different DNA methylation profiles, correlating with the expression of genes related to nervous tissue development and regeneration in patients with T1DM with autonomic neuropathy provide evidence for the role of epigenetic mechanisms promoting the development of CAN, a chronic complication of T1DM.

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Funding

Work co-financed from the scientific grant of the National Science Center no 2014/13/B/NZ4/00149.

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Authors and Affiliations

  1. University Hospital in Krakow, Kraków, Poland

    Joanna Gastoł, Łukasz Pawliński & Beata Kieć- Wilk

  2. Center for Medical Genomics- Omicron, Jagiellonian University Medical College, Kraków, Poland

    Przemysław Kapusta, Ewelina Pitera & Paweł Wołkow

  3. Department of Clinical Biochemistry, Jagiellonian University Medical College, Kraków, Poland

    Anna Polus & Maria Biela

  4. Department of Metabolic Diseases, Jagiellonian University Medical College, Kraków, Poland

    Beata Kieć- Wilk

Authors
  1. Joanna Gastoł
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  2. Przemysław Kapusta
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  4. Ewelina Pitera
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Correspondence to Beata Kieć- Wilk.

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The authors declare that they have no conflict of interest.

Ethics

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Opinion no. KBET/192/B/2014.

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Informed consent was obtained from all individual participants included in the study.

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Gastoł, J., Kapusta, P., Polus, A. et al. Epigenetic mechanism in search for the pathomechanism of diabetic neuropathy development in diabetes mellitus type 1 (T1DM). Endocrine 68, 235–240 (2020). https://doi.org/10.1007/s12020-019-02172-9

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  • DOI: https://doi.org/10.1007/s12020-019-02172-9

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